[1]刘珂杭,宗西翠,张慧丹,等.牛乳铁蛋白活性多肽LfcinB在大肠杆菌中的串联表达、纯化及生物活性分析[J].南京师范大学学报(自然科学版),2020,43(02):100-107.[doi:10.3969/j.issn.1001-4616.2020.02.016]
　Liu Kehang,Zong Xicui,Zhang Huidan,et al.Tandem Expression,Purification and Bioactivity Characterization ofBovine Lactoferricin Peptide in Escherichia coli[J].Journal of Nanjing Normal University(Natural Science Edition),2020,43(02):100-107.[doi:10.3969/j.issn.1001-4616.2020.02.016]
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牛乳铁蛋白活性多肽LfcinB在大肠杆菌中的串联表达、纯化及生物活性分析()
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《南京师范大学学报》（自然科学版）[ISSN:1001-4616/CN:32-1239/N]

卷:
第43卷
期数:
2020年02期
页码:
100-107
栏目:
·生物学·
出版日期:
2020-05-30

文章信息/Info

Title:
Tandem Expression,Purification and Bioactivity Characterization ofBovine Lactoferricin Peptide in Escherichia coli
文章编号:
1001-4616(2020)02-0100-08
作者:
刘珂杭1; 宗西翠2; 张慧丹1; 完颜杨珂1; 陈玉清1
(1.南京师范大学生命科学学院,生物化学与生物制品研究所,江苏 南京 210023)(2.南京中医药大学翰林学院,医学院西医基础教研室,江苏 泰州 225300)
Author(s):
Liu Kehang1; Zong Xicui2; Zhang Huidan1; Wanyan Yangke1; Chen Yuqing1
(1.School of Life Sciences,Nanjing Normal University,Institute of Biochemistry and Biological Products,Nanjing 210023,China)(2.Department of Basic Western Medicine,Nanjing University of Chinese Medicine Hanlin College,Taizhou 225300,China)
关键词:
Bovine lactoferricin; 抗菌肽; 串联表达; SUMO
Keywords:
Bovine lactoferricin; antimicrobial peptide; tandem expression; SUMO
分类号:
Q291
DOI:
10.3969/j.issn.1001-4616.2020.02.016
文献标志码:
A
摘要:
牛乳铁蛋白(LfcinB)是一种阳离子抗菌肽,具有抗细菌、抗肿瘤等多种生物学活性. 本文探讨了利用SUMO(Small Ubiquitin-Related Modifier)融合标签在大肠杆菌系统中通过LfcinB串联表达策略制备重组LfcinB的方法. 结果显示,大于90%的SUMO-(LfcinB)n蛋白以可溶形式表达于BL21(DE3)细胞中; SUMO-(LfcinB)2的表达量高于SUMO-LfcinB和SUMO-(LfcinB)3. 通过SUMO特异性蛋白酶切除SUMO标签,羟胺释放单体,得到纯化的重组LfcinB,产率约为15 mg/L. 生物活性结果表明,重组LfcinB具有一定的抗菌和抗肿瘤活性. 本研究为结合SUMO标签与串联表达策略大量制备获得具有生物学功能的重组抗菌肽提供了一种有效的方法.
Abstract:
Bovine lactoferricin(LfcinB)is a cationic antimicrobial peptides with activities of anti-bacteria and anti-tumor. Here it was reported that a method of producing tandem multimers of LfcinB using SUMO(small ubiquitin-related modifier)tag in Escherichia coli. Results showed that the expression level of the soluble fusion protein with SUMO tag was more than 90% of total target protein,and the dimer of LfcinB was expressed at a higher level than monomer and trimer when fused with SUMO tag. After using SUMO-specific protease to remove the SUMO tag,hydroxylamine to release monomer,the purified LfcinB was obtained with a yield of approximately 15 mg/L. The biological assays demonstrated that the recombinant LfcinB exhibited a certain antimicrobial and antitumor activity. Altogether,the SUMO fusion system with tandem repeat expression technology provides a potential production method for functional antimicrobial peptides.

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备注/Memo

备注/Memo:
收稿日期:2020-01-12.
基金项目:国家自然科学基金项目(81573337)、江苏省自然科学基金项目(BK20141446).
通讯作者:陈玉清,博士,教授,研究方向:抗菌免疫肽抗癌的分子机制. E-mail:chenyuqing@njnu.edu.cn
更新日期/Last Update: 2020-05-15